CN113679679B

CN113679679B - Preparation method of epirubicin hydrochloride for injection

Info

Publication number: CN113679679B
Application number: CN202111061300.7A
Authority: CN
Inventors: 付训忠; 张吉旺; 胡和平; 丁兆; 麻利虹; 骆第丰; 文倩; 唐棋; 嬴俊良
Original assignee: Sichuan Huiyu Haiyue Pharmaceutical Technology Co ltd; SICHUAN HUIYU PHARMACEUTICAL CO Ltd
Current assignee: Sichuan Huiyu Haiyue Pharmaceutical Technology Co ltd; SICHUAN HUIYU PHARMACEUTICAL CO Ltd
Priority date: 2021-09-10
Filing date: 2021-09-10
Publication date: 2023-04-07
Anticipated expiration: 2041-09-10
Also published as: CN113679679A

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a preparation method of epirubicin hydrochloride for injection, which comprises the steps of preserving heat of a liquid medicine at the temperature of less than or equal to 5 ℃ for 0-120 min, and then cooling to the temperature of-35 ℃ to-45 ℃ for freeze drying to obtain the epirubicin hydrochloride for injection. The method better ensures the stability of the product on the premise of ensuring that the epirubicin for injection meets the requirements of quality standards, can be widely applied to the preparation process of the freeze-dried preparation containing methylparaben, and improves the medication safety.

Description

Preparation method of epirubicin hydrochloride for injection

Technical Field

The invention relates to the technical field of pharmaceutical preparations, in particular to a preparation method of epirubicin hydrochloride for injection.

Background

The molecular formula of Epirubicin Hydrochloride (Epirubicin Hydrochloride) is C ₂₇ H ₃₀ NO ₁₁ Cl, belonging to antibiotic antitumor drugs, is a stereoisomer of adriamycin, has the characteristic of reducing cardiotoxicity, and contributes to the antitumor effect of adriamycin by targeting topoisomerase II. Epirubicin hydrochloride for injection is a sterile lyophilized powder for injection, developed by Farmitalia caro Erba, italy, first marketed in france in 1982, approved in china in 1998, approved by the FDA in 1999, and marketed in the united states in 2000. Original researchThe company Farmitalia Carlo Erba incorporated in 1995 Pharmacia&Upjohn, pfizer and Pharmacia 2003&Upjohn is combined, epirubicin hydrochloride is owned by Pfizer, and the indications comprise pancreatic cancer, esophageal cancer, multiple myeloma, breast cancer and the like.

The epirubicin hydrochloride for injection of the original research company sold in the market comprises epirubicin hydrochloride, lactose, methylparaben and water for injection, wherein the epirubicin hydrochloride is an active substance, the lactose is an excipient, the water for injection is a solvent, a freeze-dried powder preparation is obtained by removing the epirubicin hydrochloride and the lactose in the freeze-drying process, and the freeze-dried powder preparation is re-dissolved when in use.

In order to reduce the medication risk of patients, it is particularly important to ensure the clarity of epirubicin hydrochloride for injection, according to the regulations on the clarity of epirubicin hydrochloride for injection in the chinese pharmacopoeia: dissolving 5 parts of the product, 10mg each in 5ml of water, clarifying the solution; if turbidity appears, it should not be concentrated more than the standard turbidity solution No. 1 (general rule 0902 first method). Epirubicin hydrochloride for injection is originally researched and commercialized

In practical application, the clarity of the redissolved solution is found to be poor, most of the solutions are turbid solutions, the solutions cannot reach a clear state, at most, the solutions can only reach No. 1 turbidity, sometimes even are greater than No. 1 turbidity, and potential safety hazards exist. The reason is mainly that the solubility of epirubicin hydrochloride is not high and is unstable, and in the process of drying and storage, part of epirubicin hydrochloride dissociates and volatilizes HCl, so that epirubicin with low solubility exists in API, and the turbidity of the redissolved solution is increased or even exceeds the standard.

It can be seen that the existing epirubicin hydrochloride products for injection still need to be improved.

Disclosure of Invention

The invention aims to provide a freeze-drying method of a freeze-dried preparation containing methylparaben, which can improve the stability and quality of products. The invention also aims to provide a preparation method of epirubicin hydrochloride freeze-dried powder.

The inventors of the present inventionEpirubicin hydrochloride primary grinding product for injection by adopting Chinese pharmacopoeia method

(batch number: CW3848, manufacturer: pereri Wuxi pharmaceutical Co., ltd.) finds that the clarity is greater than the turbidity of No. one, and has higher risk, which indicates that the original product dissociates much epirubicin in preparation and/or storage, so that the clarity exceeds standard, and in order to better ensure the safety of patients, the product should be improved, so that the dissociation of epirubicin hydrochloride in the preparation/storage process is reduced as much as possible, and the stability of the product is improved.

Meanwhile, the inventor observes that a freeze-dried solution gradually forms a layered system in the freezing process, white loose powder appears in the upper layer, and the white loose powder which is chromatographed on the upper layer after the layering is methyl hydroxybenzoate through HPLC content detection, and the presumed reason is that the solubility of the methyl hydroxybenzoate is lower than that of epirubicin hydrochloride and lactose, and the temperature of the bottom of liquid is lower than that of the top in the freezing process, so that the methyl hydroxybenzoate moves upwards and is gathered in the upper layer to form the layering. In addition, the inventor also found that after completely freezing, a part of solid particles are carried out by the airflow in the desorption drying process due to the fluffy mass formed on the surface, and the content of methyl hydroxybenzoate in the final product is lower due to the high accumulation of methyl hydroxybenzoate on the upper layer.

In the existing literature, the methylparaben in epirubicin hydrochloride for injection is reported to be used as a bacteriostatic agent, but the epirubicin hydrochloride for injection is administered in a single dose, and the bacteriostatic agent is not required to be used theoretically, and the inventor unexpectedly discovers that if the methylparaben is not added in the prescription, the epirubicin hydrochloride freeze-dried powder is easy to be turbid after redissolving, or the redissolving turbidity is obviously increased after being placed for a period of time, but the redissolving solution can be ensured to be clear after sufficient amount of the methylparaben is added, so the methylparaben is presumed to be capable of increasing the solubility of the epirubicin hydrochloride or epirubicin so as to ensure the clarity of the solution, play a role of a solubilizer and improve the turbidity of the freeze-dried powder after being dissolved, therefore, if the content of the methylparaben in a final product can be ensured, the loss of the final product in the production process is reduced, and the stability of the epirubicin hydrochloride for injection is improved.

Therefore, a method for preparing epirubicin hydrochloride for injection is urgently needed to ensure the stable quality of epirubicin hydrochloride for injection.

To this end, the present invention provides a freeze-drying method of a freeze-dried preparation, wherein the freeze-dried preparation contains methylparaben, and the freeze-drying method comprises: (1) prefreezing: firstly, preserving heat of the liquid medicine at a first temperature for 0-120 min, and then cooling to a second temperature for heat preservation and pre-freezing; (2) sublimation drying; (3) resolving and drying; wherein the first temperature is not more than 5 ℃, and the second temperature is-35 ℃ to-45 ℃.

In the step (1), when the holding time is 0min, it should be understood that the temperature of the liquid medicine is directly reduced from the first temperature to the second temperature, and the liquid medicine is not held at the first temperature.

In the invention, the freeze-dried preparation is epirubicin hydrochloride freeze-dried powder.

The drug solution in the present invention refers to a liquid containing epirubicin hydrochloride, and may be in the form of a solution or a suspension.

In the invention, the first temperature is-40 ℃ to 5 ℃. Further, in some embodiments, the first temperature is from-5 ℃ to 5 ℃, such as-5 ℃, -4 ℃, -3 ℃, -2 ℃, -1 ℃, 0 ℃,1 ℃,2 ℃,3 ℃, 4 ℃, 5 ℃. Further, in some embodiments, the first temperature is-5 ℃.

In the invention, the pre-freezing is to pre-cool the shelf to the first temperature and then place the sample for pre-freezing, or to adjust the temperature of the shelf to the first temperature within 1-5 min after the sample is placed on the shelf and start the pre-freezing process. Further, in some embodiments, the pre-freezing procedure is initiated by adjusting the shelf temperature to the first temperature within 1min after the sample is placed on the shelf.

The expression "within 1-5 min" should be understood to include the same thing as the rest of the similar cases.

In the invention, the holding time at the first temperature is 0-80 min. Further, in some embodiments, the incubation time is 50min to 70min, such as 50min, 55min, 60min, 65min, 70min. Further, in some embodiments, the incubation time is 60min.

In the present invention, the second temperature is-40 ℃. Further, in some embodiments, the holding time at the second temperature is 60 to 180min. Further, in some embodiments, the holding time is 100min to 150min, such as 100min, 105min, 110min, 115min, 120min, 125min, 130min, 140min, 150min. Further, in some embodiments, the incubation time is 120min.

Furthermore, the time for cooling from the first temperature to the second temperature is 1-50 min.

In a specific embodiment of the present invention, the time for cooling from the first temperature to the second temperature is 1 to 40min, preferably 1min.

In the invention, the sublimation drying comprises the following steps:

a. heating from the second temperature to-33-28 ℃, and then preserving the heat for 120-480 min, wherein the heating time is 60-240 min, and the vacuum degree is kept at 0.03-0.3 mbar;

b. heating to-27-23 deg.c, maintaining the temperature for 240-480 min, heating for 120min and maintaining the vacuum degree of 0-0.07 mbar;

c. heating to-22-18 deg.c, maintaining the temperature for 240-360 min, heating for 120-600 min and maintaining the vacuum degree of 0-0.07 mbar.

Further, in some embodiments, the sublimation drying comprises the steps of:

a. heating to-30 deg.C from the second temperature, maintaining the temperature for 120min, heating for 60min, and maintaining the vacuum degree at 0.2 + -0.1 mbar;

b. heating to-25 deg.C, maintaining the temperature for 480min, heating for 120min, and maintaining the vacuum degree at 0mbar;

c. heating to-20 deg.C, maintaining the temperature for 240min, heating for 600min, and maintaining the vacuum degree at 0mbar.

In the present invention, the desorption drying includes the steps of:

i. after sublimation and drying, heating the shelf to-5 ℃, and then preserving heat for 120-180 min, wherein the heating time is 450-530 min, and the vacuum degree is kept at 0-0.22 mbar;

and ii, heating to 25-40 ℃, wherein the heating time is 120-180 min, the maintaining time is 120min, and the vacuum degree is kept at 0-0.22 mbar.

Further, in some embodiments, the desorption drying comprises the steps of:

i. after sublimation drying, heating the shelf to 0 deg.C, maintaining the temperature for 180min, wherein the heating time is 480min, and the vacuum degree is maintained at 0mbar;

and ii, heating to 25 ℃, wherein the heating time is 180min, the maintaining time is 120min, and the vacuum degree is kept at 0mbar.

The epirubicin hydrochloride freeze-dried powder comprises methylparaben, and is freeze-dried by adopting any one of the freeze-drying methods; the preparation method comprises the following steps:

I. mixing the raw materials;

II, filtering: pre-filtering the mixed liquid medicine and then performing sterilization filtration;

III, filling;

performing freeze-drying by using the freeze-drying method in any one of the above steps;

further, in some embodiments, the pre-filtering is: pre-filtering the medicinal liquid with a filter made of 0.45 μm +0.22 μm PVDF material; the degerming filtration comprises the following steps: filtering the pre-filtered liquid medicine by 2 filters made of PVDF with the diameter of 0.22 μm and the diameter of 0.22 μm.

Further, in some embodiments, the sterilizing filtration is performed under a protective gas atmosphere.

The raw material mixing, filtering and filling are all common process steps in the field, and the preparation method which is formed by adopting any one or more of the steps and the freeze drying process belongs to the protection scope of the invention.

The raw material mixing refers to mixing the raw materials in the preparation formula.

The protective gas is a gas which is inert to the epirubicin or epirubicin hydrochloride reaction, is used for isolating the epirubicin or epirubicin hydrochloride from oxygen, and does not chemically react with the epirubicin or other components in the sterilization process, and the optional protective gas includes but is not limited to nitrogen, helium, neon, argon, krypton or xenon.

In the embodiment of the invention, the protective gas is nitrogen.

Further, the filling container may be any container in the art that can withstand the freeze-drying process of the present invention, including but not limited to injection bottles, vials, ampoule bottles, polyethylene bottles, and cartridge bottles.

Further, in some embodiments, the raw materials are mixed as follows: mixing and dissolving the auxiliary materials and water with the amount less than the formula amount to obtain a liquid medicine A, mixing epirubicin hydrochloride with the liquid medicine A to obtain a liquid medicine B, and mixing the residual amount of water with the liquid medicine B to obtain a liquid medicine C;

further, the water is water for injection;

further, the total amount of water used is 250mL per gram of epirubicin hydrochloride.

In a specific embodiment of the invention, the epirubicin hydrochloride freeze-dried powder comprises the following components: 240 to 580 parts of epirubicin hydrochloride, 1500 to 2500 parts of lactose and 50 to 100 parts of methyl hydroxybenzoate;

further, the epirubicin hydrochloride freeze-dried powder comprises the following components: 400 parts of epirubicin hydrochloride, 2104 parts of lactose and 80 parts of methyl hydroxybenzoate.

The invention has the beneficial effects that:

(1) The freeze-drying method of the freeze-dried preparation containing methylparaben provided by the invention ensures the quality and stability of the freeze-dried preparation. Experiments show that the method better ensures the stability of the product on the premise of ensuring that the freeze-dried preparation meets the requirements of quality standards, can be widely applied to the preparation process of the freeze-dried preparation containing methylparaben, and improves the medication safety.

(2) The epirubicin hydrochloride freeze-dried powder prepared by the freeze-drying method can improve the stability of epirubicin hydrochloride in the product, reduce the dissociation degree of the epirubicin hydrochloride in the preparation/storage process and simultaneously ensure that the content of methylparaben is not lost. Through detection, the indexes of related substances and the like of the product prepared by the invention are superior to those of imported products

In the accelerated stability test, the data of the stability test show that the product is stable when stored at 60 ℃ and that the total impurity content is much lower than that of the inlet product after 30 days>

The total impurity content is higher, and the quality is better.

Detailed Description

The invention provides a preparation method of epirubicin hydrochloride for injection. Those skilled in the art can modify the process parameters appropriately in view of the disclosure herein. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the methods and applications of this invention have been described in terms of preferred embodiments, it will be apparent to those of ordinary skill in the art that variations and modifications in the methods and applications described herein, as well as other suitable variations and combinations, may be made to implement and use the techniques of this invention without departing from the spirit and scope of the invention. The test materials adopted by the invention are all common commercial products and can be purchased in the market. The invention is further illustrated by the following examples:

example 1

Prescription:

TABLE 1

The preparation method of epirubicin hydrochloride for injection comprises the following steps:

step 1: to a 1L beaker was added about 70% of the formulation volume of purified water, the stirring was turned on, and the mixture was heated to 40-50 ℃ at a speed of 100-150rpm.

Step 2: stirring a prescribed amount of methylparaben in a container with about 2% of water for injection (60-80 ℃) for 2min each time, standing for 10s, pouring the suspension into a beaker, and continuously stirring for 5 times at a rotating speed of 100-300rpm;

and 3, step 3: adding lactose in a prescription amount into a beaker, using 2% of water for injection to trickle and wash a container containing the lactose, and putting the washing liquid into the medicinal liquid, wherein the rotating speed is set to be 100-300rpm;

and 4, step 4: adding the API into the beaker, and continuously stirring to completely dissolve the API and the auxiliary materials;

and 5: adding purified water to the total weight, cooling to less than or equal to 30 ℃, and continuously stirring for more than or equal to 10min;

and 6: filtering the liquid medicine obtained in the step 5 through a sterilizing filter head with the diameter of 0.22 mu m;

and 7: starting the filling machine, and adjusting the speed and the filling amount; filling the liquid medicine obtained in the step 6 into a 10ml borosilicate glass tube injection bottle, wherein the filling speed is less than or equal to 450 bottles/min;

and 8: and (3) setting freeze drying parameters according to table 2, precooling the shelf to-40 ℃, closing the box door after the sample filled in the step (7) is put into the box, and starting a freeze drying program. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 2 summary of lyophilization process parameters

Example 2

Prescription:

TABLE 3

Composition of	Dosage of
		Epirubicin hydrochloride	4g
Lactose	21.04g
		Hydroxy phenyl methyl ester	800mg
Water for injection	In sufficient amount, add up to 1L
		Is made into	400 bottle (2.5 ml/bottle)

step 1: adding purified water with about 70 percent of preparation volume into a 1L beaker, starting stirring, heating to 40-50 ℃, and setting the rotating speed at 100-150rpm;

and 2, step: stirring the prescribed amount of methyl hydroxybenzoate in a container with about 2% water for injection (60-80 deg.C) for 2min each time, standing for 10s, pouring the suspension into a beaker, continuously stirring for 5 times at 100-300rpm;

and step 3: adding lactose in a prescription amount into a beaker, using 2% of water for injection to trickle and wash a container containing the lactose, and putting the washing liquid into the medicinal liquid, wherein the rotating speed is set to be 100-300rpm;

step 6: filtering the liquid medicine obtained in the step 5 through a sterilizing filter head with the diameter of 0.22 mu m;

and 8: and (3) setting freeze drying parameters according to table 4, precooling the shelf to 5 ℃, closing the box door after the sample filled in the step (7) is put into the box, and starting a freeze drying program. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 4 summary of lyophilization process parameters

Example 3

Prescription:

TABLE 5

Composition of	Dosage of
		Epirubicin hydrochloride	400g
Lactose	2104g
		Hydroxy phenyl methyl ester	80g
Water for injection	In sufficient amount, adding up to 100L
		Is made into	40000 bottle (2.5 ml/bottle)

step 1: adding about 70 percent of injection water with the preparation volume into a 500L liquid preparation tank, cooling to 40-50 ℃, starting stirring, and setting the rotating speed to be 100-150rpm;

step 2: stirring the methyl hydroxybenzoate with the prescription amount by using about 2% of water for injection (60-80 ℃) for 2min in a container, standing for 10s, pouring suspension in the container into a liquid preparation tank, continuously performing the step for 5 times, rinsing the container, and continuously stirring at the rotating speed of 100-300rpm;

and step 3: adding lactose of a prescribed amount into a liquid preparation tank, using 2% of water for injection to trickle and wash a container containing lactose, and setting the rotating speed of the washing liquid in the liquid medicine to be 100-300rpm;

and 4, step 4: the epirubicin hydrochloride with the prescription dose is slowly added into a stainless steel barrel containing injection water (40-50 ℃) with the preparation volume of 5 percent in a feeding isolator, pre-dissolved and then added into a liquid preparation tank, the container containing the epirubicin hydrochloride is flushed with about 2 percent of the preparation dose of the injection water (40-50 ℃) for 2 times each time, and the lotion is added into the liquid medicine. Stirring for more than or equal to 10min to completely dissolve the API and the auxiliary materials;

and 5: adding water for injection to total weight, cooling to less than or equal to 30 deg.C, and stirring for more than or equal to 10min;

and 6: filtration

(1) Pre-filtering: pressurizing the liquid preparation tank by using clean nitrogen, controlling the pressure to be 1.0-2.0 Bar, and filtering the liquid medicine into the liquid storage tank through a 0.45-0.22-micron prefilter;

(2) And (3) degerming and filtering: and introducing nitrogen into the liquid storage tank, and controlling the pressure to be 1.0-2.0 bar. Filtering the liquid medicine into a filling buffer tank through 2 paths of 0.22 mu m +0.22 mu m sterilizing filters, and continuing the step until the filling is finished. When the liquid medicine is stored in the liquid storage tank, recording the temperature and the filtering pressure of the liquid medicine every 30min for 1 time;

and 7: starting the filling machine, and adjusting the speed and the filling amount; filling the liquid medicine obtained in the step 6 into a 10ml borosilicate glass tube injection bottle, wherein the filling speed is less than or equal to 450 bottles/minute;

and 8: and (4) setting freeze drying parameters according to table 6, closing the box door after the filling of the sample in the step (7) is finished, and starting a freeze drying program. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 6 summary of lyophilization process parameters

Comparative example 1

Prescription:

TABLE 7

Composition of	Dosage of
		Epirubicin hydrochloride	4g
Lactose	21.04g
		Water for injection	In sufficient quantity, add up to 1L
Is made into	400 bottle (2.5 ml/bottle)

step 1: adding about 70% of purified water with the preparation volume into a 1L beaker, starting stirring, heating to 40-50 ℃, and setting the rotating speed at 100-150rpm;

step 2: adding lactose in a prescription amount into a beaker, using 2% of injection water to trickle and wash a container containing the lactose, and setting the rotation speed of the washing liquid in the medicinal liquid to be 100-300rpm;

and step 3: adding the API, and continuously stirring to completely dissolve the API and the auxiliary materials;

and 4, step 4: adding purified water to the total weight, cooling to less than or equal to 30 ℃, and continuously stirring for more than or equal to 10min;

and 5: filtering the liquid medicine obtained in the step 4 by a sterilizing filter head with the diameter of 0.22 mu m;

and 6: starting the filling machine, and adjusting the speed and the loading amount; filling the liquid medicine obtained in the step 5 into a 10ml borosilicate glass tube injection bottle, wherein the filling speed is less than or equal to 450 bottles/minute;

and 7: and (4) setting freeze drying parameters according to the table 8, closing the box door after the filled sample in the step 6 is put into the box, and starting a freeze drying program. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 8 summary of lyophilization process parameters

Comparative example 2

Prescription:

TABLE 9

Composition of	Dosage of
		Epirubicin hydrochloride	4g
Lactose	21.04g
		Hydroxy benzene methyl ester	800mg
Water for injection	In sufficient quantity, add up to 1L
		Is made into	400 bottle (2.5 ml/bottle)

and step 3: adding lactose in a prescription amount into a beaker, using 2% of injection water to trickle and wash a container containing the lactose, and setting the rotation speed of the washing liquid in the medicinal liquid to be 100-300rpm;

and 7: starting the filling machine, and adjusting the speed and the loading amount; filling the liquid medicine obtained in the step 6 into a 10ml borosilicate glass tube injection bottle, wherein the filling speed is less than or equal to 450 bottles/minute;

and 8: and (3) setting freeze drying parameters according to the table 10, precooling the shelf to-40 ℃, closing the box door after the sample filled in the step (7) is put into the box, and starting a freeze drying program. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 10 summary of lyophilization process parameters

Phases	Set temperature (. Degree. C.)	Setting time (min)	Maintenance time (min)	Set vacuum degree (mbar)
					Shelf pre-cooling	-40	NA	NA	NA
Prefreezing	-40	1	120	NA
					One time sublimation	-5	120	360	0.2±0.02
Drying by desorption	5	60	360	0.2±0.02

Comparative example 3

Prescription:

TABLE 11

Composition of	Amount of the composition
		Epirubicin hydrochloride	4g
Lactose	21.04g
		Hydroxy benzene methyl ester	800mg
Water for injection	In sufficient quantity, add up to 1L
		Is made into	400 bottle (2.5 ml/bottle)

and 2, step: stirring a prescribed amount of methylparaben in a container with about 2% of water for injection (60-80 ℃) for 2min each time, standing for 10s, pouring the suspension into a beaker, and continuously stirring for 5 times at a rotating speed of 100-300rpm;

and 8: and (3) setting freeze drying parameters according to table 12, precooling the shelf to-40 ℃, closing the box door after the sample filled in the step (7) is put into the box, and starting a freeze drying program. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 12 summary of freeze-drying process parameters

Comparative example 4

Prescription:

watch 13

Composition of	Dosage of
		Epirubicin hydrochloride	200g
Lactose	1052g
		Hydroxy benzene methyl ester	40g
Water for injection	In sufficient amount, add up to 50L
		Is made into	20000 bottles (2.5 ml/bottle)

step 1: about 70% of the formulation volume of water for injection was added to a 500L dispensing tank and cooled to 40-50 ℃. Starting stirring, and setting the rotating speed at 100-150rpm;

step 2: stirring the prescribed amount of methylparaben in a container with about 2 percent of water for injection (60-80 ℃) for 2min each time, standing for 10s, pouring the suspension into a liquid preparation tank, continuously performing the step for 5 times, rinsing the container, and continuously stirring at the rotating speed of 100-300rpm;

and 3, step 3: adding lactose in a prescription amount into the liquid preparation tank, using 2% of water for injection to trickle and wash a container containing the lactose, and setting the rotating speed of the washing liquid to be 100-300rpm when the washing liquid is added into the liquid medicine;

and 4, step 4: the epirubicin hydrochloride with the prescription amount is slowly added into a stainless steel barrel containing injection water (40-50 ℃) with the preparation volume of 5 percent in a feeding isolator, pre-dissolved and then added into a liquid preparation tank, the container containing the epirubicin hydrochloride is flushed with the injection water (40-50 ℃) with the preparation amount of about 2 percent each time for 2 times, and the lotion is added into the liquid medicine. Stirring for more than or equal to 10min to completely dissolve the API and the auxiliary materials;

step 6: and (3) filtering:

(1) And (3) filtering: pressurizing the liquid preparation tank by using clean nitrogen, controlling the pressure to be 1.0-2.0 Bar, and filtering the liquid medicine into the liquid storage tank through a 0.45 mu m +0.22 mu m prefilter;

and step 8: freeze drying parameters were set according to table 14, and after the shelf was pre-cooled to 5 ℃, the filled sample of step 7 was put into the box and the box door was closed to start the freeze drying procedure. And after freeze-drying, re-pressurizing to 800mbar with nitrogen, pressing a plug, taking out of a box, and rolling a cover.

Table 14 summary of lyophilization process parameters

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And (3) product quality detection:

1. detection of related substances

High performance liquid chromatography is adopted to detect the epirubicin for injection and imported products obtained in examples 1 and 3 and comparative examples 1, 2 and 4

Wherein the chromatographic detection conditions are shown in table 15:

TABLE 15 chromatographic detection conditions of epirubicin hydrochloride related substances for injection

The results of the detection of the relevant substances are shown in Table 16:

TABLE 16 epirubicin hydrochloride for injection and the results thereof

2. Detection of content of methylparaben

The content of methylparaben in the epirubicin for injection obtained in examples 1 to 3 and comparative examples 2 to 4 was determined by high performance liquid chromatography, wherein the chromatographic conditions are shown in table 17:

TABLE 17 chromatographic detection conditions for content of epirubicin hydrochloride

Chromatographic column	Welch Ultimate LP-C18 4.6×250mm，5μm
		Detecting wavelength	272nm
Column temperature	35℃
		Flow rate of flow	1.3ml/min
Sample size	10μl
		Temperature of sample chamber	5℃
Run time	37min
		Diluent	6.8g/L Potassium dihydrogen phosphate-methanol (40
Mobile phase	6.8g/L Potassium dihydrogen phosphate-methanol (40
		Time of acquisition	20min

The results of the measurement of the methylparaben content are shown in table 18:

TABLE 18 relative content test results of epirubicin hydrochloride for injection (based on the prescribed amount as 100%)

Note: the intermediate refers to the liquid before freeze-drying, i.e. the liquid state of the final freeze-dried powder.

According to the results in table 18, the content of methylparaben in the epirubicin hydrochloride freeze-dried powder obtained by the preparation method can reach more than 98% of the formula amount, and the content of methylparaben in the product prepared in example 3 can reach 100%, so that the methylparaben is not lost in the drying process, and the product quality is improved; the sublimation drying process of the comparative example 3 is different, the analysis drying temperature of the comparative example 4 is higher, and the content of methyl hydroxybenzoate in the finally obtained product is obviously lower.

3. Physical and chemical property detection

(1) The characteristics are as follows: the product was taken and observed visually in bright light.

(2) Clarity: 1 bottle of the product is taken, and water is added according to the marked amount to prepare a solution containing 2mg in each 1 ml. The samples were placed in a pair of turbidimetric glass tubes, respectively, in comparison with the turbidity standard solution No. 1, and prepared for 5 minutes. The illumination intensity is 1000Lux when the umbrella is placed under the umbrella shed lamp, and the observation and comparison are carried out in the direction vertical to the black background plate.

(3) Moisture content: measuring 1ml of formamide solution, adding the formamide solution into a coulometer for measurement, measuring 3 parts in parallel, continuously calibrating for 3 times to obtain a result RSD (reference signal density) less than or equal to 4%, and recording the measured water as B.

Taking 1 bottle of the test article, removing the label and the aluminum cover, wiping the outer wall of the container with ethanol, drying, and precisely weighing and marking as Ma.

2ml of formamide are added to the sample by means of a syringe, wiped off and weighed again, and this is recorded as Mb. Shaking thoroughly to dissolve the contents completely, and optionally performing ultrasonic treatment to confirm complete dissolution of the sample without insoluble substances.

Another clean syringe is taken out to extract 1ml of the dissolving solution, and the dissolving solution is directly injected into a coulometric moisture tester for measurement, and the water content is recorded as A.

After the measurement, the content of the vessel was poured out, the vessel was washed with water, dried and precisely weighed, and designated as Mc.

A: the water content of the test solution%

B: blank solvent water content%

And Ma: total weight of sample tape packaging material, g

Mb: total weight of dissolved sample solution tape wrapping material, g

Mc: total weight of packing material, g.

The results of the physical and chemical property measurements are shown in Table 19:

TABLE 19 physicochemical Properties test results for epirubicin hydrochloride for injection

According to the results in Table 19, the clarity of the French New product of the original product is poor by the detection of the method of Chinese pharmacopoeia; comparative example 1 no methylparaben was added, and after 30 days of storage at 60 ℃, the turbidity of the product increased, indicating that epirubicin hydrochloride partially dissociates, there is a risk of medication, and the longer the storage time, the greater the risk, sufficient methylparaben should be added to ensure the stability of the product; although the content of methylparaben in comparative example 2 was also about 100%, it was found from table 19 that the water content was high, which was not favorable for long-term storage; the product prepared by the embodiment of the invention can still have good clarity after long-time high-temperature storage, which shows that the product prepared by the invention has better redissolution stability, can reduce the dissociation degree of epirubicin hydrochloride in the preparation/storage process and ensures that the content of methylparaben is not lost, thereby reducing the medication risk, and the product has lower water content and is more favorable for ensuring the stability of the product.

4. Stability test

The epirubicin for injection obtained in experimental examples 1-2 and comparative examples 1, 2 and 4 and imported products

The mixture is placed at 60 ℃ for 30 days, and the content of the relevant substances is respectively considered.

The results of the measurement of the contents of the relevant substances are shown in Table 20:

TABLE 20 influence factors of epirubicin hydrochloride for injection (60 deg.C, 30 days) on the results of the detection of the related substances

In the table, impurity A is doxorubicin ketone, impurity C is doxorubicin, and impurity G is an epirubicin dimer. As can be seen from Table 20, the indexes of epirubicin hydrochloride for injection, such as each impurity and the total impurity content, prepared by the method of the invention are superior to those of imported products

And according to the results, the content of methylparaben in the freeze-dried product obtained in example 3 can be maintained at 100%. In accelerated stability experiments, the data from the stability experiments in example 3 show that the product is storage-stable at 60 ℃ and remains clear after reconstitution and that the total impurity content is much lower than that of the inlet product after 30 days>

The total impurity content shows that the quality and the stability of the epirubicin hydrochloride for injection prepared by the method are superior to those of the original research reagent->

The foregoing is only a preferred embodiment of the present invention, and it should be noted that it is obvious to those skilled in the art that various modifications and improvements can be made without departing from the principle of the present invention, and these modifications and improvements should also be considered as the protection scope of the present invention.

Claims

1. A freeze-drying method of a freeze-dried preparation is characterized in that the freeze-dried preparation is epirubicin hydrochloride freeze-dried powder; the epirubicin hydrochloride freeze-dried powder comprises the following components: 400 parts of epirubicin hydrochloride, 2104 parts of lactose and 80 parts of methyl hydroxybenzoate, wherein 250mL of water is used per gram of epirubicin hydrochloride;

the freeze-drying method comprises the following steps: after the sample is placed on a shelf, adjusting the temperature of the shelf to a first temperature within 1min, and starting a pre-freezing procedure;

(1) Pre-freezing: firstly, preserving heat of the liquid medicine for 60min at a first temperature, and then cooling to a second temperature for heat preservation and pre-freezing; (2) sublimation drying; (3) resolving and drying; wherein the first temperature is-5 ℃ and the second temperature is-40 ℃; the time for cooling from the first temperature to the second temperature is 1min, and the heat preservation time at the second temperature is 120min;

the sublimation drying comprises the following steps:

c. heating to-20 deg.C, maintaining the temperature for 240min, heating for 600min, and maintaining vacuum degree of 0mbar;

the desorption drying comprises the following steps:

i. after sublimation and drying, heating the shelf to 0 deg.C, and maintaining the temperature for 180min, wherein the heating time is 480min, and the vacuum degree is kept at 0mbar;

2. A freeze-drying method of a freeze-dried preparation is characterized in that the freeze-dried preparation is epirubicin hydrochloride freeze-dried powder; the epirubicin hydrochloride freeze-dried powder comprises the following components: 400 parts of epirubicin hydrochloride, 2104 parts of lactose and 80 parts of methyl hydroxybenzoate, wherein 250mL of water is used per gram of epirubicin hydrochloride;

the freeze-drying method comprises the following steps: pre-cooling the shelf to 5 ℃, placing the sample on the shelf, and starting a pre-freezing program;

(1) Pre-freezing: cooling the medicinal liquid to-40 deg.C, and keeping the temperature for 120min, wherein the cooling time is 40min;

(2) Sublimation drying, comprising the following steps:

a. heating to-30 deg.C from pre-freezing temperature, maintaining the temperature for 120min, heating for 60min, and maintaining vacuum degree of 0.2 + -0.1 mbar;

c. heating to-20 deg.C, maintaining the temperature for 240min, heating for 600min, and maintaining the vacuum degree at 0mbar;

(3) The analytical drying comprises the following steps:

i. after sublimation drying, heating the shelf to 0 deg.C, maintaining the temperature for 120min, wherein the heating time is 480min, and the vacuum degree is maintained at 0mbar;

and ii, heating to 25 ℃, then preserving the heat for 120min, wherein the heating time is 120min, and the vacuum degree is kept at 0mbar.

3. A method for preparing epirubicin hydrochloride freeze-dried powder, which is characterized in that the epirubicin hydrochloride freeze-dried powder comprises methylparaben, and is freeze-dried by the freeze-drying method of any one of claims 1-2;

the epirubicin hydrochloride freeze-dried powder comprises the following components: 400 parts of epirubicin hydrochloride, 2104 parts of lactose and 80 parts of methyl hydroxybenzoate, wherein 250mL of water is used per gram of epirubicin hydrochloride;

the preparation method comprises the following steps:

I. mixing the raw materials;

III, filling;

lyophilizing by the lyophilization method of any one of claims 1-2;

the pre-filtering is as follows: pre-filtering the liquid medicine with a filter membrane or a filter element of 0.45 mu m +0.22 mu m;

the degerming filtration comprises the following steps: filtering the liquid medicine obtained by pre-filtering for 1-3 times by a filter membrane or a filter element with the diameter of 0.22 mu m +0.22 mu m;

the sterilization filtration is carried out under the protective gas atmosphere.

4. The method of claim 3, wherein the pre-filtering is: pre-filtering the liquid medicine by a PVDF material filter with the diameter of 0.45 mu m +0.22 mu m; the degerming filtration comprises the following steps: filtering the pre-filtered liquid medicine for 2 times by a PVDF material filter with the diameter of 0.22 mu m +0.22 mu m; the sterile filtration was performed under nitrogen atmosphere.

5. The method according to claim 3, wherein the raw materials are mixed as follows: mixing and dissolving the auxiliary materials and water with the amount less than the formula amount to obtain a liquid medicine A, mixing epirubicin hydrochloride with the liquid medicine A to obtain a liquid medicine B, and mixing the residual amount of water with the liquid medicine B to obtain a liquid medicine C;

the water is water for injection.